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Title:Three-dimensional reconstructions from cryoelectron microscopy images reveal an intimate complex between helicase DnaB and its loading partner DnaC.
Authors:C. San Martin
Sample:DnaB.DnaC complex from E.coli
Aggregation state:Single particle (42.2 angstroms resolution)
Red flagLatest update:2011-05-26
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bullet_white.gif  Summary
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bullet.png Experimental details
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bullet_white.gif  Visualization
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Sample
Sample name: DnaB.DnaC complex from E.coli
Oligomeric state: one homohexamer of DnaB binds to one homohexamer of DnaC
Theoretical molecular weight of the sample: 0.48
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinDnaBhomohexamertrueEscherichia coliGO:0003678IPR001198
2proteinDnaC6 molecules of DnaC bound to a DnaB hexamertrueEscherichia coliGO:0000166IPR003593
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.6.04 mg/mL50 mM Tris.HCl pH 7.6 2 mM DTT 5 mM MgCl2 200 mM NaCl 0.25 mM ADP
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGERsamples were adsorbed onto carbon-coated molybdenum holey grids after 30s glow discharge, and vitrified by quick plunging in liquid ethane in a double-side blotting device msVitrification instrument: double side blotting device
Imaging:
MicroscopeVoltageIllumination modeImaging modeCs:Defocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS EM420100 kVFLOOD BEAMBRIGHT FIELD2 mm1500 nm2000 nm60000LAB6Kodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Gatan626GATAN LIQUID NITROGEN°45° eV98 K K K mrad10 e/Å2
Processing
Software:Xmipp, SPIDER
Resolution by author:42.2 Å
Resolution method:DPR at 45 degrees
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
18 μm/pixel8linkEIKONIX IEEE 488
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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